SureGuide CRISPR Libraries GeCKOv2 Catalog Libraries Benefits SureGuide CRISPR Libraries SureGuide Catalog CRISPR Libraries Agilent’s pooled libraries come in a variety of formats designed to suit your unique experimental requirements. Pooled libraries require both uniform representation and high fidelity to reduce screening, improve analysis and speed your time to results. All of our libraries, whether standard, genome-wide content or fully customized pools, are synthesized on our industry-leading DNA manufacturing system, so you can be sure of getting the same quality and consistency out of every assay. Our CRISPR libraries are also one of the few licensed solutions for conducting research with pooled guide libraries, making sure that you have not only the best libraries available but the freedom to use them for the research you’ve designed. – Exome-wide knock-down screening Validated guides targeting all known genes – Highest quality, sequence-verified GeCKOv2 libraries – Lentiviral, ready-to-package plasmid libraries – Human and mouse libraries - Common model organisms/cell lines for genomewide screening – Full flexibility to develop your research applications Catalog Ready-to-Package Plasmid Library Agilent’s catalog CRISPR guide libraries target all exonic sequences in the human and mouse genome and are ideal for genome-wide functional screening. These libraries contain content developed by and licensed by a well known research institute. In comparison to other GeCKO libraries, Agilent’s GeCKO libraries contain fewer missed guides and more uniform representation across the library (Table 1). Both the mouse and human GeCKO libraries come as plasmids, with a puromycin selection marker and a GFP reporter, that can be used to generate high-titers of lentiviral particles. Integration of SureGuide CRISPR library in Genomics Assay Workflow OLS Library PCR amplify Clone into viral vector Transform E. coli host Isolate plasmid DNA Transfect packaging line Catalog Ready-to-Package Library Custom Ready-to-Clone Library Custom Ready-to-Amplify Library Figure 1. Agilent offers CRISPR libraries in a variety of formats, from plasmid libraries to unamplified oligo pools. Isolate viral supernatant GeCKO Libraries Guide Representation Comparison with Competitor Libraries Species Human Mouse Genes targeted 19,050 20,611 6 6 1,864 1,175 4 4 Targeting constructs per gene miRNA targeted Targeting constructs per miRNA Control (nontargeting) sgRNA Total sgRNA constructs Viral plasmid vector 1,000 1,000 123,411 130,209 Single and dual vector.lentiCRISPRv2 and lenitGuide-Puro Missed Guides 90th/10th percentile 95th/5th percentile 99.5th/0.5th percentile Agilent option 1 1 2.32 3.04 6.72 Agilent option 2 1 2.47 3.38 11.06 Agilent option 3 1 2.38 3.19 9.83 Agilent option 4 1 1.99 2.64 8.30 GeCKO (Broad) ? 8.73 16.00 NA GeCKO (Competitor) 39 5.29 9.83 68.40 GeCKO (Competitor) expanded 204 6.00 11.95 333.00 Table 2. With <.01% missed guides, you can be sure the guides in your library will be present in your delivered library. The uniformity of our libraries has an average 90/10 ratio of 2.29, which is less than half that of our competitor’s libraries (5.29). Table 1. The GeCKOv2 library contains experimentally validated CRISPR guides targeting all human or mouse genes. Instructions to order GeCKOv2 Libraries GeCKOv2 libraries are available for both mouse (G7554A) and human (G7555A) applications. Table 1 provides a summary of the libraries including the number of guides per gene, non-coding targets, and control sequences. Kits also include a control appropriate for the organism the library is designed for. If you require additional information, including the sequences of each component of the library please contact your local Agilent representative or technical support. Ordering Information SureGuide GeCKOv2 CRISPR Libraries SureGuide Amplified Custom CRISPR Libraries SureGuide Unamplified Custom CRISPR Libraries Part Numbers G7553A - Human G7554A - Mouse G7555A - Human (hU6 promoter) G7555B - Any species Library Format Ready-to-package plasmid library Ready-to-clone amplified DNA library Ready-to-amplify DNA library Delivery Lentivirus Designed for lentivirus, user choice Users choice Number of Guides Human - 123,411; Mouse - 130,209 Up to 60,000 Up to 100,000 User Supplies Lentivirus packaging, delivery, screening Cloning, viral packaging, delivery screening Amplification, cloning, viral packaging, delivery, screening Sequence Verified Yes No No Quantity 200 ug of plasmid DNA split into A and B libraries 125 ng of linear, amplified, pooled library 10 pmol of linear, unamplified, pooled library Also Included N/A N/A Amplification kit Custom Design Input None Up to 50 nt sequence for each guide Complete sequence up to 200 nt Control Mal1-C User designed User designed www.agilent.com/genomics/CRISPR For Research Use Only. Not for use in diagnostic procedures. This information is subject to change without notice. PR7000-0245 © Agilent Technologies, Inc. 2017 Published in the USA, September, 2017 5991-7347EN